Pe anti human hla a2 antibody

The candidate peptides were synthesized in GenScript Biotechnology Co., Ltd (Nanjing, China) and resuspended in DMSO at a concentration of 10 mM, respectively. T2A2 cells were seeded into 96-well plates, and then incubated with peptides at a final concentration of 20 μM at 37°C for 4 hours. Set DMSO as blank control, Influenza A M1 peptide (GILGFVFTL) as positive control, and Zika virus peptide (GLQRLGYVL) as negative control. Cells were stained with PE anti-human HLA-A2 antibody (BioLegend) at 4°C in the dark for 30 min, and acquired in flow cytometer FACS Canto (BD).

Antibody were purchased from BioLegend, including FITC anti-human HLA-A2 antibody (Cat#343303), PE anti-human HLA-A2 antibody (Cat#343305), anti-human CD28 antibodies (Cat#302901), APC anti-human-CD69 (Cat#310909), APC anti-human CD8 (Cat#344721), PerCP anti-human IFN-γ (Cat#502524), and FITC anti-human granzyme B (Cat#515403). Flex-T monomer (Cat#280003) and PE conjugated streptavidin (Cat#405203) were purchased from BioLegend. Ficoll-Paque PLUS (Cat#17144003) was from GE and DMSO (Cat#D2650) was from Sigma-Aldrich. IMDM (Cat#SH30228.01) and penicillin-streptomycin (Cat#SV30010) were from HyClone, and FBS (Cat#S711-001S) was purchased from LONSERA. PBS (Cat#C10010500BT) was from Gibco (Waltham, Massachusetts, USA). D-biotin (Cat#2110450) was purchased from Invitrogen (Waltham, Massachusetts, USA), and Mitomycin C (Cat#50-07-7) was from Sinochem Holdings (Beijing, China). CFSE (Cat#T6802) was from Targetmol (Boston, Massachusetts, USA). EasySep Human CD8⁺ T Cell Isolation Kit (Cat#17953) was from Stem Cell Technologies (Vancouver, British Columbia, Canada), and IL-2 (recombinant human interleukin-2(¹²⁵Ala) injection) was from SL PHARM (Beijing, China). Leuko act cktl with GolgiPlug (Cat#550583) was purchased from BD.